The air-gap PAD: a roll-to-roll-compatible fabrication method for paper microfluidics.

Paper-based analytical devices (PADs) offer a low-cost, user-friendly platform for rapid point-of-use testing. Without scalable fabrication methods, however, few PADs make it out of the academic laboratory and into the hands of end users. Previously, wax printing was considered an ideal PAD fabrication method, but given that wax printers are no longer commercially available, alternatives are needed. Here, we present one such alternative: the air-gap PAD. Air-gap PADs consist of hydrophilic paper test zones, separated by "air gaps" and affixed to a hydrophobic backing with double-sided adhesive. The primary appeal of this design is its compatibility with roll-to-roll equipment for large-scale manufacturing. In this study, we examine design considerations for air-gap PADs, compare the performance of wax-printed and air-gap PADs, and report on a pilot-scale roll-to-roll production run of air-gap PADs in partnership with a commercial test-strip manufacturer. Air-gap devices performed comparably to their wax-printed counterparts in Washburn flow experiments, a paper-based titration, and a 12-lane pharmaceutical screening device. Using roll-to-roll manufacturing, we produced 2700 feet of air-gap PADs for as little as $0.03 per PAD.

Activation/proliferation and apoptosis of bystander goat lymphocytes induced by a macrophage-tropic chimeric caprine arthritis encephalitis virus expressing SIV Nef.

Caprine arthritis encephalitis virus (CAEV) is the natural lentivirus of goats, well known for its tropism for macrophages and its inability to cause infection in lymphocytes. The viral genome lacks nef, tat, vpu and vpx coding sequences. To test the hypothesis that when nef is expressed by the viral genome, the virus became toxic for lymphocytes during replication in macrophages, we inserted the SIVsmm PBj14 nef coding sequences into the genome of CAEV thereby generating CAEV-nef. This recombinant virus is not infectious for lymphocytes but is fully replication competent in goat macrophages in which it constitutively expresses the SIV Nef. We found that goat lymphocytes cocultured with CAEV-nef-infected macrophages became activated, showing increased expression of the interleukin-2 receptor (IL-2R). Activation correlated with increased proliferation of the cells. Interestingly, a dual effect in terms of apoptosis regulation was observed in exposed goat lymphocytes. Nef was found first to induce a protection of lymphocytes from apoptosis during the first few days following exposure to infected macrophages, but later it induced increased apoptosis in the activated lymphocytes. This new recombinant virus provides a model to study the functions of Nef in the context of infection of macrophages, but in absence of infection of T lymphocytes and brings new insights into the biological effects of Nef on lymphocytes.

Simian immunodeficiency virus Vpr/Vpx proteins kill bystander noninfected CD4+ T-lymphocytes by induction of apoptosis.

The depletion of CD4+ T-lymphocytes central to the immunodeficiency in acquired immunodeficiency syndrome (AIDS) is largely mediated by apoptosis of both infected and uninfected cells, but the mechanisms involved and the viral proteins responsible are still poorly characterized. It has recently been suggested that, in human and simian immunodeficiency virus (HIV) and SIV, Vpr is a major modulator of apoptosis in infected cells. Recently, we have reported on a chimera of caprine arthritis-encephalitis virus (CAEV) carrying vpr/vpx genes from SIVmac239, which is replication competent in goat macrophages but not in lymphocytes or human cells. Despite infection being restricted to macrophages, inoculation of primary goat peripheral blood mononuclear cells (PBMCs) with this chimera induced apoptosis in the lymphocyte population. In addition, when infected goat synovial membrane (GSM) cells were co-cultured with human CD4+ T lymphocyte SupT1 cell line, these CD4+ T cells showed increased apoptosis. The parental CAEV induced no significant apoptosis in goat PBMC cultures or in co-cultures with human SupT1 lymphocytes. This indicates that SIV Vpr/Vpx proteins indeed mediate apoptosis of T-lymphocytes and, moreover, do so without the need for active infection of these cells. Moreover, this apoptosis was observed when SupT1s were cocultured in direct contact, but not in absence of contact with CAEV-pBSCAvpxvpr-infected GSM cells. In view of these data, we propose that SIV Vpx/Vpr activate cell-to-cell contact-dependent extracellular signaling pathways to promote apoptotic death of uninfected bystander T-lymphocytes. Understanding this mechanism might bring insight for intervening in the loss of CD4+ T lymphocytes in the SIV infection model and in human AIDS.

Composition and bioactivity of the leaf essential oil of Heteropyxis dehniae from Zimbabwe.

The leaf oil of Heteropyxis dehniae Suess. (Heteropyxidaceae) was obtained by hydrodistillation and analyzed by GC/MS. The most abundant essential oil components are linalool (58.3%), 4-terpineol (9.8%), alpha-terpineol (3.6%), and caryophyllene oxide (3.1%). The antimicrobial activity against Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Candida albicans, and Aspergillus niger, and the in vitro cytotoxicity of the oil on PC-3, MDA-MB-231, Hs 578T, MCF7, SK-MEL-28, and 5637 human tumor cells were also examined. Caryophyllene oxide shows notable cytotoxic activity with LC50 values of 147-351 microM.

Cytotoxic activity of Ozoroa insignis from Zimbabwe.

The crude methanol bark extract of the Zimbabwean medicinal plant, Ozoroa insignis, showed in-vitro cytotoxic activity against Hep-G2 (human hepatocellular carcinoma), MDA-MB-231 (human mammary adenocarcinoma), and 5637 (human primary bladder carcinoma). Bioactivity-directed chromatographic separation led to isolation of anacardic acid and ginkgoic acid as the cytotoxic components.